Streptonigrin is a potent, although toxic, anticancer agent produced by Streptomyces flocculus. The work proposed herein will continue our studies on the biosynthesis of streptonigrin and our recently initiated project to develop a biomimetic synthesis of streptonigrin and streptonigrin analogs. The apparently new pathway leading to quinoline rings that we have discovered this past year using UL-13C6-D-glucose will be followed up with feeding experiments to determine the exact nature of the bio-intermediates. We will also investigate the stereochemistry of the beta-methylation of tryptophan that occurs early in the pathway. This will provide additional insights to the scope of the chemistry of biological systems, and may provide additional opportunities for mutasynthetic approaches to streptonigrin analogs. Our discovery of the tryptophan-derived origin of the phenylpicolinic acid portion of streptonigrin and the mode of cleavage of the beta-carboline intermediate provides an opportunity to develop a rather straightforward biomimetic synthesis. We are currently working on the synthesis of 10' desmethoxy streptonigrin and 9', 10'-bisdesmethoxy-11-methoxy streptonigrin, and hope to begin the synthesis of streptonigrin itself.